Open chromatin assays are known to have significant GC bias. Please take this
into consideration as necessary.
Library complexity quality metrics
Library complexity (filtered non-mito BAM)
rep1
rep2
rep3
ctl1
Total Fragments
3081465
994220
1873536
2630100
Distinct Fragments
2960864
966479
1809428
2565235
Positions with Two Read
98183
24370
55640
49343
NRF = Distinct/Total
0.960862
0.972098
0.965782
0.975337
PBC1 = OneRead/Distinct
0.96419
0.97322
0.967182
0.978896
PBC2 = OneRead/TwoRead
29.076683
38.596512
31.453037
50.890643
Mitochondrial reads are filtered out by default.
The non-redundant fraction (NRF) is the fraction of non-redundant mapped reads
in a dataset; it is the ratio between the number of positions in the genome
that uniquely mapped reads map to and the total number of uniquely mappable
reads. The NRF should be > 0.8. The PBC1 is the ratio of genomic locations
with EXACTLY one read pair over the genomic locations with AT LEAST one read
pair. PBC1 is the primary measure, and the PBC1 should be close to 1.
Provisionally 0-0.5 is severe bottlenecking, 0.5-0.8 is moderate bottlenecking,
0.8-0.9 is mild bottlenecking, and 0.9-1.0 is no bottlenecking. The PBC2 is
the ratio of genomic locations with EXACTLY one read pair over the genomic
locations with EXACTLY two read pairs. The PBC2 should be significantly
greater than 1. See more details at
the ENCODE portal standard for ChIP-Seq pipeline
NRF (non redundant fraction)
PBC1 (PCR Bottleneck coefficient 1)
PBC2 (PCR Bottleneck coefficient 2)
PBC1 is the primary measure. Provisionally
0-0.5 is severe bottlenecking
0.5-0.8 is moderate bottlenecking
0.8-0.9 is mild bottlenecking
0.9-1.0 is no bottlenecking
Replication quality metrics
IDR (Irreproducible Discovery Rate) plots
Reproducibility QC and peak detection statistics
overlap
idr
Nt
6645
1619
N1
6698
412
N2
2059
762
N3
8558
1414
Np
2488
1118
N optimal
6645
1619
N conservative
6645
1619
Optimal Set
rep2_vs_rep3
rep2_vs_rep3
Conservative Set
rep2_vs_rep3
rep2_vs_rep3
Rescue Ratio
2.6708199356913185
1.4481216457960644
Self Consistency Ratio
4.156386595434677
3.4320388349514563
Reproducibility Test
fail
borderline
Reproducibility QC
N1: Replicate 1 self-consistent peaks (comparing two pseudoreplicates generated by subsampling Rep1 reads)
N2: Replicate 2 self-consistent peaks (comparing two pseudoreplicates generated by subsampling Rep2 reads)
Ni: Replicate i self-consistent peaks (comparing two pseudoreplicates generated by subsampling RepX reads)
Nt: True Replicate consistent peaks (comparing true replicates Rep1 vs Rep2)
Np: Pooled-pseudoreplicate consistent peaks (comparing two pseudoreplicates generated by subsampling pooled reads from Rep1 and Rep2)
Self-consistency Ratio: max(N1,N2) / min (N1,N2)
Rescue Ratio: max(Np,Nt) / min (Np,Nt)
Reproducibility Test: If Self-consistency Ratio >2 AND Rescue Ratio > 2, then 'Fail' else 'Pass'
Number of raw peaks
rep1
rep2
rep3
Number of peaks
11451
30631
32314
Top 300000 raw peaks from spp with FDR 0.01
Peak calling statistics
Peak region size
rep1
rep2
rep3
idr_opt
overlap_opt
Min size
75.0
60.0
85.0
72.0
72.0
25 percentile
300.0
240.0
340.0
290.0
290.0
50 percentile (median)
300.0
240.0
340.0
290.0
290.0
75 percentile
300.0
240.0
340.0
290.0
290.0
Max size
300.0
240.0
340.0
290.0
290.0
Mean
299.79993013710595
239.77581535046195
339.26483876957354
277.23038912909203
286.8887885628292
Enrichment / Signal-to-noise ratio
Strand cross-correlation measures (trimmed/filtered SE BAM)
rep1
rep2
rep3
Number of Subsampled Reads
3081465
994220
1873536
Estimated Fragment Length
110
115
135
Cross-correlation at Estimated Fragment Length
0.543254666937325
0.309445130222828
0.439852166972061
Phantom Peak
40
40
40
Cross-correlation at Phantom Peak
0.5380755
0.2936322
0.4205608
Argmin of Cross-correlation
1500
1500
1500
Minimum of Cross-correlation
0.5296676
0.2720251
0.4028237
NSC (Normalized Strand Cross-correlation coeff.)
1.025652
1.137561
1.091922
RSC (Relative Strand Cross-correlation coeff.)
1.615991
1.731837
2.087625
Performed on subsampled (15000000) reads mapped from FASTQs that are trimmed to 50.
Such FASTQ trimming and subsampling reads are for cross-corrleation analysis only.
Untrimmed FASTQs are used for all the other analyses.
NOTE1: For SE datasets, reads from replicates are randomly subsampled to 15000000.
NOTE2: For PE datasets, the first end (R1) of each read-pair is selected and trimmed to 50 the reads are then randomly subsampled to 15000000.
Normalized strand cross-correlation coefficient (NSC) = col9 in outFile
Relative strand cross-correlation coefficient (RSC) = col10 in outFile
Estimated fragment length = col3 in outFile, take the top value
Jensen-Shannon distance (filtered/deduped BAM)
rep1
rep2
rep3
AUC
0.36930057424542123
0.3150714906728927
0.32450650626550687
Synthetic AUC
0.4935016495738625
0.48861667147850274
0.4917156096925494
X-intercept
0.04024280530692769
0.0652993076099077
0.04772724099204679
Synthetic X-intercept
7.841135688659982e-203
2.7531429241172766e-65
2.9160889261399934e-124
Elbow Point
0.49294943018948467
0.5424241033937288
0.5946834973379586
Synthetic Elbow Point
0.5026314531094559
0.4990357187344033
0.5127055171540771
JS Distance
0.03678385223857956
0.10774506411403753
0.09937735400807503
Synthetic JS Distance
0.1738920623606129
0.22952751518540712
0.23654053595318375
% Genome Enriched
38.65076689533559
35.23728972465974
36.56721330442224
Diff. Enrichment
7.668555225800711
17.372685187068104
17.564356347039467
CHANCE Divergence
0.06521454412951344
0.14799813166266887
0.14962250750703707
Peak enrichment
Fraction of reads in peaks (FRiP)
FRiP for spp raw peaks
rep1
rep2
rep3
rep1-pr1
rep2-pr1
rep3-pr1
rep1-pr2
rep2-pr2
rep3-pr2
pooled
pooled-pr1
pooled-pr2
Fraction of Reads in Peaks
0.07534891236290925
0.21836936878425842
0.24537007141489042
0.2328162765176762
0.14139531431375502
0.26777792978903875
0.23068812991052493
0.1459233662125161
0.2686042630038033
0.4355815255372418
0.07133564538868396
0.07943584165536008
FRiP for overlap peaks
rep1_vs_rep2
rep1_vs_rep3
rep2_vs_rep3
rep1-pr1_vs_rep1-pr2
rep2-pr1_vs_rep2-pr2
rep3-pr1_vs_rep3-pr2
pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks
0.040288472990851115
0.040449706632787956
0.062209140144454944
0.04871745672678798
0.05270732368607973
0.1080449831723534
0.04163208667365808
FRiP for IDR peaks
rep1_vs_rep2
rep1_vs_rep3
rep2_vs_rep3
rep1-pr1_vs_rep1-pr2
rep2-pr1_vs_rep2-pr2
rep3-pr1_vs_rep3-pr2
pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks
0.022392925691922038
0.021970770941259463
0.034056532675731235
0.009902849871745672
0.033660234820626855
0.05076859933783895
0.02785579887195396
For spp raw peaks:
repX: Peak from true replicate X
repX-prY: Peak from Yth pseudoreplicates from replicate X
pooled: Peak from pooled true replicates (pool of rep1, rep2, ...)
pooled-pr1: Peak from 1st pooled pseudo replicate (pool of rep1-pr1, rep2-pr1, ...)
pooled-pr2: Peak from 2nd pooled pseudo replicate (pool of rep1-pr2, rep2-pr2, ...)
For overlap/IDR peaks:
repX_vs_repY: Comparing two peaks from true replicates X and Y
repX-pr1_vs_repX-pr2: Comparing two peaks from both pseudoreplicates from replicate X
pooled-pr1_vs_pooled-pr2: Comparing two peaks from 1st and 2nd pooled pseudo replicates